Subcellular Scaled Protein Binding Template by Dip-Pen Nanolithography and its Application to Single Cell Patterning

Micro- and nano-patterns of protein are importantly demanded in fields of biotechnology, such as tissue engineering, biosensors, biodiagnostics, and cell-biology, because a highly integrated biochip with high sensitivity essentially needs small volume of capture proteins (ex. antibody or DNA) on a substrate. Another attractive application of protein patterning is cell engineering. Cell binding and patterning have been recently reported by subcellular scaled extracellular materials (ECM) patterns. Here, we demonstrate new method of subcellular scaled protein patterning using lithographically fabricated silane template on silicon oxide surface. Subcellular scaled IgG protein arrays were obtained by binding on dip-pen nanolithography (DPN) printed aldehyde terminated silane (aldehyde-silane) micro- and nano-array templates on silicon oxide surface. Remarkably, the aldehyde–silane template shows durable reactivity with IgG proteins even until 1 year. Moreover, subcellular scaled ECM patterns, fibronectin micro- and nano-arrays, were obtained by binding on the DPN printed aldehyde-silane template. Finally, single cell patterning was also demonstrated using subcellular scaled fibronectin patterns based upon the DPN printed aldehyde-silane template. In summary, we demonstrated that subcellular scaled protein patterning and single cell patterning on silicon oxide substrate are available by DPN printed aldehyde-silane template